deleting a certain gene from bacterium - (Nov/16/2009 )
pDNA on Nov 22 2009, 08:50 AM said:
If you want to delete a section of the genome completely you will have to use the "scarless engineering" approach published by Feher et al.. If you are interested read it up or if something is unclear to you ask again!
Regards,
p
Regards,
p
I cant acces that file
@fishdoc and Homebrew, at the moment I can not sequence it, so I have to deal with it as it is.
-lyok-
lyok on Nov 23 2009, 09:29 AM said:
@fishdoc and Homebrew, at the moment I can not sequence it, so I have to deal with it as it is.
So you're going to attempt to make a mutant using a plasmid of unknown sequence? How will you know that you've actually made the correct mutation then? And in the event you do get a mutation made, you will have to sequence that to verify the correct sequence was manipulated. Based on my experience with bacterial mutagenesis, I will recommend strongly against going any further without verifying that plasmid sequence. If you don't know the sequence of the plasmid, you won't know what it does to the sequence of the genome. Without a genotype, any resulting phenotype is pretty worthless, because you can't make any direct correlation to the genotype you think you have. In other words, there is no shot of getting anything published, patented, or whatever you want to do with sequencing that plasmid. If you are by chance in a graduate program, there's no way that should even get by your committee. That's a major snag, in other words.
-fishdoc-
fishdoc on Nov 23 2009, 05:23 PM said:
lyok on Nov 23 2009, 09:29 AM said:
@fishdoc and Homebrew, at the moment I can not sequence it, so I have to deal with it as it is.
So you're going to attempt to make a mutant using a plasmid of unknown sequence? How will you know that you've actually made the correct mutation then? And in the event you do get a mutation made, you will have to sequence that to verify the correct sequence was manipulated. Based on my experience with bacterial mutagenesis, I will recommend strongly against going any further without verifying that plasmid sequence. If you don't know the sequence of the plasmid, you won't know what it does to the sequence of the genome. Without a genotype, any resulting phenotype is pretty worthless, because you can't make any direct correlation to the genotype you think you have. In other words, there is no shot of getting anything published, patented, or whatever you want to do with sequencing that plasmid. If you are by chance in a graduate program, there's no way that should even get by your committee. That's a major snag, in other words.
You are 100% correct.
But at the moment I have to work with it as it is. The plasmid is being sequenced, so for the future thats no problem.
But at the moment we are working with it as it is.
thanks for the help and pDNA thank you for the paper.
-lyok-